Ammonia-free synthesis of 3-trifluoromethyl-3-phenyldiaziridine

<p>An ammonia-free synthesis of 3-trifluoromethyl-3-phenyldiaziridine, an important intermediate in the synthesis of a widely used photolabel 3-trifluoromethyl-3-phenyldiazirine, is described. By avoiding the use of volatile, corrosive, and toxic anhydrous ammonia, the major hazard involved in the synthesis of this widely used photolabel is eliminated. Furthermore, this synthesis is convenient compared to the conventional route, since it is significantly less time consuming and, due to the absence of liquid ammonia, this method does not require the maintenance of low temperature for prolonged periods.</p

Metal-Free Arylation of Ethyl Acetoacetate with Hypervalent Diaryliodonium Salts: An Immediate Access to Diverse 3‑Aryl-4(1<i>H</i>)‑Quinolones

A clean arylation protocol of ethyl acetoacetate was developed using hypervalent diaryliodonium salts under mild and metal-free conditions. The scope of the reaction, using symmetric and unsymmetric iodonium salts with varying sterics and electronics, was examined. Further, this method has been applied for the synthesis of antimalarial compound <b>ELQ-300</b>, which is currently in preclinical development

Protocol for Isolation of Labeled Proteome from Neuroblastoma Cell Lysate using Probe 1.

<p>Cell lysate sample treated with probe 1 under UV was loaded on monomeric avidin column and washed with PBS to elute non-bound proteome, 2mM biotin to elute weakly bound proteome and 2.5M glycine (pH = 2.8) to elute strongly bound proteome.</p

Gene Ontology (GO) molecular function categories observed using BiNGO Analysis for the PBS Wash.

<p>The nodes represent various molecular functions identified by this analysis where color represents significance of enrichment (orange, yellow, white represent order of significance). The arrows represent association of the broad functional categories (larger circles) and related subcategories (smaller circles).</p

Top 10 statistically over-represented GO Biological Process terms in Overall Identified Proteins, according to BiNGO.

<p>Top 10 statistically over-represented GO Biological Process terms in Overall Identified Proteins, according to BiNGO.</p

ABPP Photoaffinity Probes.

<p>Structure of the current probe <b>1</b> differs from earlier reported probes based on the labeling functionality in the 8-position versus the 5’-position.</p

Synthesis and Evaluation of a Novel Adenosine-Ribose Probe for Global-Scale Profiling of Nucleoside and Nucleotide-Binding Proteins

<div><p>Proteomics is a powerful approach used for investigating the complex molecular mechanisms of disease pathogenesis and progression. An important challenge in modern protein profiling approaches involves targeting of specific protein activities in order to identify altered molecular processes associated with disease pathophysiology. Adenosine-binding proteins represent an important subset of the proteome where aberrant expression or activity changes of these proteins have been implicated in numerous human diseases. Herein, we describe an affinity-based approach for the enrichment of adenosine-binding proteins from a complex cell proteome. A novel <i>N</i>⁶-biotinylated-8-azido-adenosine probe (AdoR probe) was synthesized, which contains a reactive group that forms a covalent bond with the target proteins, as well as a biotin tag for affinity enrichment using avidin chromatography. Probe specificity was confirmed with protein standards prior to further evaluation in a complex protein mixture consisting of a lysate derived from mouse neuroblastoma N₁₈TG₂ cells. Protein identification and relative quantitation using mass spectrometry allowed for the identification of small variations in abundance of nucleoside- and nucleotide-binding proteins in these samples where a significant enrichment of AdoR-binding proteins in the labeled proteome from the neuroblastoma cells was observed. The results from this study demonstrate the utility of this method to enrich for nucleoside- and nucleotide-binding proteins in a complex protein mixture, pointing towards a unique set of proteins that can be examined in the context of further understanding mechanisms of disease, or fundamental biological processes in general.</p></div

Gene Ontology (GO) molecular function categories observed using BiNGO Analysis for the Regeneration Wash.

<p>The nodes represent various molecular functions identified by this analysis where color represents significance of enrichment (orange, yellow, white represent order of significance). The arrows represent association of the broad functional categories (larger circles) and related subcategories (smaller circles).</p

Synthesis of AdoR Photoaffinity Probe 1.

<p>Reaction conditions: a) Ac₂O, DMAP, dry DCM, 0°C, RT, 8 h, 95%; b) aq. Br₂, 10% Na₂HPO₄, dioxane, RT, 6–7 d, 92%; c) dry DCM, POCl₃, <i>N</i>, <i>N</i>-diethylaniline, reflux, 10 min, 78%; d) mono-<i>N</i>-Boc-1,6-diaminohexane, dry EtOH, -10°C, 12 h, 66%; e) CsN_3, TMSN₃, dioxane/H₂O, 3 d, 79%; f) TFA, DCM, RT, 4 h, 81%; g) biotin, EDCI, DMAP, dry DCM, RT, 8 h, 72%; h) NH₃, MeOH, -40°C, 2 h, preparative HPLC, 61%.</p

Top 10 statistically over-represented GO Biological Process terms in PBS Wash, according to BiNGO.

<p>Top 10 statistically over-represented GO Biological Process terms in PBS Wash, according to BiNGO.</p